牛膝多糖对CD4+T细胞的诱导和分化作用研究

目的：探讨牛膝多糖免疫调节作用的机制。 方法： (1)以牛膝多糖 (800 mg/L)分别对哮喘和肺癌病人的(PBMC)进行体外诱导培养，在18 h以RT-PCR检测IFN-γ和IL-4的基因表达率。(2)用牛膝多糖在不同时间内和不同剂量分别对Th细胞进行体外诱导其分化，并分别收集细胞悬液和抽提细胞中RNA，分别采用RT-PCR和ELISA检测Th细胞所分泌的IFN-γ和IL-4的含量。 结果： (1)哮喘和肺癌患者PBMC中IFN-γ mRNA表达率分别由6/25升到14/25(P<0.01)、3/22升到10/22(P<0.05)，IL-4 mRNA的表达率由17/25降到9/25(P<0.05)、14/22降到5/22(P<0.05)。 (2)Th细胞培养至6 h，即可检到IFN-γ mRNA分泌，18 h达到高峰，之后迅速下降，48 h已难以检出。而IL-4被抑制。(3) IFN-γ蛋白表达量存在时间依赖性，从18-24 h时段开始，蛋白表达量开始显著增加(P<0.05)，但在24 h、36 h和48 h的3个时段内，蛋白表达量增加不明显(P>0.05)，进入平台期。IL-4蛋白表达量被抑制。(4)IFN-γ基因水平和蛋白表达量分别与牛膝多糖的诱导浓度相关(r=0.979)。400 mg/L和800 mg/L是较好的的诱导浓度。而IL-4的蛋白表达被抑制。 结论： (1)应用牛膝多糖能初步纠正肺癌和哮喘患者Th1和Th2细胞因子的失平衡。(2)牛膝多糖能在转录水平和翻译水平促进Th1类细胞因子的分泌，而抑制Th2类细胞因子的分泌。     

肺癌患者外周血单个核细胞Th1/Th2反应状态及黄芪的调节作用

目的:探讨肺癌患者外周血单个核细胞（PBMC）Th1/Th2免疫反应状态,观察中药黄芪（AG）对其作用。方法:采集并分离肺癌患者和健康志愿者的PBMCs,各分为4组:其中1组即刻冻存待测,其余3组分别为不加药对照组、加 10％AG、20％AG组,培养48h,收集细胞。以IL-2、IFN-γ代表Th1型细胞因子,IL-4、IL-6、IL-10代表Th2型细胞因子,用RT-PCR方法检测肺癌患者PBMC中Th1/Th2型细胞因子mRNA的表达。结果:肺癌患者PBMC中,IL-4、IL-6和IL-10表达阳性率显著高于正常对照（P<0.05）,而IL-2无1例表达（0/23）,IFN-γ仅1例表达;经与黄芪培养后（2种浓度）,肺癌患者Th2型因子与未加AG培养组相比,IL-4、IL-6和IL-10的表达率下降（P<0.05）;而IL-2的表达率显著提高（P<0.05）。结论:肺癌患者外周血的免疫细胞呈现Th2型免疫反应优势状态;黄芪对于肺癌宿主PBMCs Th1/Th2的状态有良好的调节作用,可使肺癌患者PBMCs的Th2型优势免疫反应向Th1方向逆转。     

膜表达的HLA-G1对同种反应性PBMC分泌Th1/Th2型细胞因子的影响

目的研究人脐静脉内皮细胞系(ECV304)表达的HLA-G1对同种异体外周血单个核细胞(PBMC)Th1/Th2型细胞因子分泌的影响。方法采用脂质体介导的基因转染技术将pcDNA3-HLA-G1转入ECV304,以间接免疫荧光技术在蛋白质水平上检测HLA-G1分子在ECV304上的表达;以表达HLA-G1的ECV304作为刺激细胞,灭活后,与健康人PBMC共同培养,用ELISA检测上清液中Th1/Th2型细胞因子的浓度,观察HLA-G1对同种异体抗原激活的PBMC分泌Th1/Th2型细胞因子的影响。结果与转染pcDNA3空质粒的对照组相比,HLA-G1能使PBMC的IL-10分泌增加(P<0.05),而IL-2,IFN-γI、L-4分泌无明显影响。结论HLA-G1能引起由同种异体抗原激活的PBMC分泌IL-10增加,提示HLA-G1有可能使Th1/Th2型细胞因子向Th2型偏移。     

中药多糖对柔嫩艾美尔球虫感染鸡细胞因子水平的影响

本文选用天然免疫活性多糖．香菇（Lentinus edodes）、银耳（Tremella fuifformisBerk）和黄芪（Astragalus membranaceus Bge）多糖，研究其对柔嫩艾美尔球虫感染鸡细胞因子IFN-γ和IL-2水平的影响。选择180只雏鸡并将其随机分为9组：3个多糖提取物添加组（LenE、TreE和AstE），3个添加提取物并免疫接种疫苗组（LenE＋V、TreE＋V和AstE＋V），1个球虫免疫组和2个对照组（球虫感染组和非感染组）。检测球虫感染后第7、14天鸡血清IFN-γ效价和脾淋巴细胞IL-2的水平。结果显示，球虫感染后第7、14天，添加多糖提取物并疫苗免疫组的血清IFN-γ效价显著高于单纯疫苗组（P〈0．01）。然而，单纯提取物添加组与单纯疫苗组之间没有显著差异。3种多糖提取物相比，感染第7天后，AstE组的IFN-γ效价最高，而TreE＋V组的IFN-γ的效价显著高于LenE＋v组和AstE＋v组。脾脏细胞IL-2产量与血清IFN-γ效价的表现基本一致。感染后第7天，提取物加疫苗免疫组的平均IL．2水平显著（P〈0．01）高于单纯疫苗组，提取物添加组的平均IL-2水平与单纯疫苗组之间没有显著差异。感染第14天后，多糖提取物添加组及多糖提取物加疫苗组的平均IL-2水平都与单纯疫苗组没有显著差异。IL-2产量在不同多糖提取物添加组间没有显著差异。本实验结果表明，多糖提取物抗球虫作用可能与刺激免疫细胞分泌IFN-γ和IL-2等细胞因子、提高T-细胞免疫应答有关；中药免疫活性多糖对球虫感染鸡有很好的免疫保护作用，当多糖与疫苗一起使用时，效果尤为明显。     

棘球蚴病患者血清中可溶性PD-1/PD-L1与相关细胞因子的水平变化

目的检测包虫患者血清中可溶性程序性死亡分子1(sPD-1)及其配体sPD-L1,Th1型细胞因子IFN-γ、Th2型细胞因子IL-6以及Th17细胞因子IL-17的分泌水平,探讨它们在包虫病感染过程中的相关作用。方法采用细胞因子磁珠阵列测定(CBA)法检测包虫患者血清中Th1、Th2、Th17细胞因子的分泌水平;ELISA检测可溶性PD-1、PD-L1的表达水平。结果与健康对照组相比,棘球蚴病患者血清中可溶性PD-1的浓度略升高但低于sPD-L1,sPD-L1浓度升高明显,Th2、Th17细胞相关细胞因子的分泌水平明显升高(P0.01),而Th1细胞相关细胞因子分泌水平未见明显变化(P0.05)。结论棘球蚴病感染过程中存在Th1/Th2的表达失衡,而炎性细胞Th17细胞参与机体免疫调节。sPD-1与sPD-L1通过动态平衡参与机体免疫调控,促进棘球蚴免疫逃逸。     

喘可治注射液对人外周血单个核细胞Th1/Th2细胞因子谱的影响

目的：通过研究喘可治注射液对人外周血单个核细胞（PBMCs）Th1／Th2细胞因子谱的影响，探讨喘可治注射液的免疫调节作用机制。方法：以流式微球分析（CBA）法检测不同处理情况下，人外周血单个核细胞分泌Th1（IFN-γTNF-α、IL-2）和Th2（IL4、IL-6、IL-10）细胞因子水平。结果：健康人PBMCs体外培养12小时后，上清中细胞因子主要为TNF-α和IL-6，喘可治使Th1和Th2细胞因子全面升高；喘可治对PDB加离子霉素诱导的PBMCs分泌Th1和Th2细胞因子具有抑制作用，并能抑制流感病人异常升高的INF-γ、TNF-α、IL-6和IL-10分泌。结论：喘可治注射液上调健康人PBMCs分泌TH1和Th2细胞因子，对异常活化的PBMCs分泌的Th1和Th2细胞因子则具有下调作用。     

内毒素对慢性HBV感染者Th1/Th2类细胞因子的影响

目的 通过研究慢性HBV感染者血浆内毒素(Endotoxin,ET)与Th1、Th2类细胞因子的关系,进而探讨其各自对慢性乙型肝炎(CHB)患者免疫功能的影响.方法 无菌收集HBV感染者和正常对照者血浆、血清,以鲎试剂法测定血浆ET水平,同时测定血清补体活性及Th1、Th2类细胞因子水平.分析ET与Th1、Th2类细胞因子的关系.结果 CHB患者IL-4与IL-10水平明显升高,IL-2、IL-12和IFN-γ水平明显降低;ET阳性组患者IL-4与IL-10水平明显高于阴性组,IL-12水平在阳性组明显低于阴性组;ET与AP50呈显著正相关.结论 CHB患者均有不同程度的Th1/Th2类细胞因子失衡,主要表现为:Th1类细胞因子减少而Th2类细胞因子增加,ET主要影响Th2类细胞因子,进而影响患者的免疫功能.     

调节性T细胞能有效降低抗CD3单克隆抗体OKT3引起的人外周血单个核细胞细胞因子的释放

目的 探索细胞因子风暴因子体外模型的构建方法及调节性T细胞(Treg)的调节作用。方法 培养健康成人外周血单核细胞(PBMC),随机分为PBMC组、对照组(PBS处理)、 CD3单克隆抗体OKT3处理组、 OKT3联合Treg处理组。处理48 h后，收集上清液，采用ELISA检测血清转化生长因子β(TGF-β)、细胞毒性T淋巴细胞相关蛋白4(CTLA4)、 γ干扰素(IFN-γ)、白细胞介素10(IL-10)、 IL-2、 IL-7和IL-6水平。结果 与对照组相比，OKT3能明显增加PBMC的TGF-β、 CTLA4、 IL-10、 IL-2、 IL-7、 IFN-γ和IL-6水平。Treg能有效减少OKT3引起的上述细胞因子释放。结论 成功构建了细胞因子风暴细胞模型，Treg能降低PBMC的细胞因子释放。     

强肌健力方对脾肾两虚大鼠细胞因子水平的影响

目的：观察强肌健力方对实验性脾肾两虚证大鼠血清细胞因子含量变化的影响,探讨脾肾两虚证与细胞因子的关系,以期进一步了解该方药对改善脾肾两虚证的意义及黄芪在方中的作用。方法：采用随机法将72只SD大鼠分为五组,正常对照组10只,脾肾两虚组16只,强肌健力方组15只,黄芪减量组15只,黄芪组16只。造模分两个阶段完成。第1d～14d采用大黄造模;第15d～24d采用氢化可的松复制脾肾两虚证大鼠模型。应用大黄及氢化可的松造模,分别给予蒸馏水、强肌健力方、黄芪减量方,黄芪方灌胃,连续给药24d后,采用放射免疫分析观察各组动物血清IL-2、IL-6、TNF水平的变化。结果：脾肾两虚组大鼠血清IL-2、TNF含量比正常对照组低（P〈0.05～0.01）,而IL-6含量则升高（P〈0.01）。强肌健力方组能升高IL-2、TNF含量和降低IL-6含量（P〈0.01）。黄芪减量组能使TNF含量升高及降低IL-6含量。黄芪组能使IL-6含量降低,但对IL-2、TNF含量没有影响。结论：脾肾两虚证的发生与细胞因子网络调节系统的失衡有关,强肌健力方可以改善脾肾两虚证大鼠这种免疫功能失衡,通过健脾益气功效来调节细胞因子水平,从而达到增强免疫功能的作用。这为脾肾两虚证时体内细胞因子水平的改变提供了理论依据,同时也提示该方重用黄芪可能是防治脾肾两虚的一个关键。     

支气管哮喘患儿治疗前后外周血Th1／Th2相关细胞因子的临床分析

目的：探讨支气管哮喘患儿治疗前后外周血Th1／Th2相关细胞因子水平的变化及临床意义。方法i应用放射免疫分析和酶联法对31例支气管哮喘患儿进行了外周血中Th1型细胞因子（IL-2、IFN-1）和Th2型细胞因子IL-4、IL-10浓度,并与35名正常人组作比较。结果i支气管哮喘患儿在治疗前血清IL-2、IFN-^y水平均非常显著地低于正常儿组（P〈0．01）,而血清IL4、IL-10水平又非常显著地高于正常儿组（P〈0．01）。经治疗1个月后则与正常儿组比较无显著性差异（P〉0．05）。结论：支气管哮喘患儿存在Th1／Th2细胞免疫失衡。Th1免疫功能低下。IFN-1下调可能是参与哮喘发病机制的重要原因,对哮喘有负向调节作用。哮喘患儿存在Th2细胞免疫功能亢进,以IL-4、IL-10为代表的Th2免疫功能在儿童哮喘的发病中发挥正性促炎作用。     

The effect of treatment with montelukast on in vitro interleukin-10 production of mononuclear cells of children with asthma

BACKGROUND: The effects of leukotriene modifiers on IL-10 production have not been studied in children with asthma. OBJECTIVE: The primary objective of this study was to determine the changes in IL-10 concentrations, clinical efficacy and peripheral blood eosinophil counts after treatment with montelukast. METHODS: The study was conducted on 27 patients: 13 patients monoallergic to grass pollen during the pollen season (GPs group) and out of the pollen season (GPos group), and on 14 patients monoallergic to house dust mite (HDM) from May to September (HDM group). Main outcome measures were changes in concentrations of IL-10 in the supernatant after a 4-week treatment with montelukast. Measurements of asthma severity score, forced expiratory volume in 1 s (FEV1) and peripheral blood eosinophil counts were the secondary end-points. RESULTS: Montelukast resulted in a within-group significant increase in IL-10 concentration in the supernatant in the GPs (54.0 vs. 125.5 pg/mL) and in the HDM (51.2 vs. 77.1 pg/mL) group. Montelukast had no effect on changes of IL-10 concentration in the supernatant from peripheral blood mononuclear cell (PBMC) culture after non-sensitizing allergen stimulation. Montelukast significantly improved asthma control and FEV1, and significantly decreased eosinophil blood count in the GPs and in the HDM group after a 4-week treatment. Montelukast did not lead to changes of all measured parameters within the GPos group. CONCLUSION: Montelukast increased IL-10 concentration in supernatants from sensitizing allergen-stimulated PBMC culture obtained from children with asthma monoallergic to grass pollen during the pollen season, and from children with asthma monoallergic to HDM.     

Profiles of immunoregulatory cytokine production in vitro in patients with IgA nephropathy and their kindred.

We hypothesized that the altered immunoglobulin synthesis and/or lymphocyte function apparent in patients with IgA nephropathy (IgAN) is due to a primary defect in lymphokine regulation. In addition, we reasoned that such changes in lymphokine production might be, at least partially, genetically determined. To assess the extent of lymphocyte abnormalities, we investigated the profile of cytokine production from peripheral blood mononuclear cells (PBMC) in 34 IgAN patients and 44 of their first degree relatives, 10 of whom had persistent microhaematuria. Compared with healthy volunteers (n = 34), PBMC from patients showed increased IL-2 production both spontaneously or after phytohaemagglutinin (PHA) (20 micrograms/ml) stimulation, whereas IL-4 and interferon-gamma (IFN-gamma) production were significantly higher only after stimulation. Microhaematuric relatives had a similar pattern of cytokine production, whereas non-microhaematuric relatives showed no significant difference versus normals. The altered pattern of cytokine production appeared to be quite specific to IgAN patients and their microhaematuric relatives, because patients with other forms of primary glomerulonephritis (n = 17) did not differ from normal individuals. Patients and relatives that hyperproduced IL-4 were also hyperproducers of IL-2. No such congruence was seen in any other group or with any other pairing of cytokines. We propose that a subpopulation of IgAN patients bear lymphocytes intrinsically hyperresponsive. Among those individuals such hyperresponsiveness may be causally related to the pathogenesis and/or character of IgAN.     

艾叶挥发油治疗大鼠变应性鼻炎的实验研究

目的:观察艾叶挥发油对变应性鼻炎(AR)大鼠血清中IgE、IL-4和IL-5含量的影响。方法:60只SD大鼠按体重随机分为正常对照组、模型组、治疗组和阳性药对照组,每组10只。实验组用卵蛋白(OVA)、氢氧化铝和灭活百日咳杆菌皮下注射致敏,用10%的卵蛋白生理盐水溶液滴鼻激发,建立AR模型。正常对照组以等量生理盐水代替。治疗组和阳性药对照组分别灌胃给予艾叶挥发油和氯雷他定治疗10天,正常对照组用橄榄油代替药物;另外20只大鼠用于被动皮肤过敏原试验(PCA),检验大鼠模型制备及治疗情况;用ELISA法测定各组大鼠外周血清IL-4、IL-5和IgE含量。结果:模型组大鼠鼻喷嚏平均次数明显多于正常对照组、治疗组和阳性对照组(P<0.05);模型组大鼠PCA试验阳性率为92%,其余三组全部表现为阴性;模型组IgE均显著高于正常对照组(P<0.05);治疗组IgE、IL-4、IL-5均显著低于模型组(P<0.05);治疗组各项指标与阳性对照组比较差异无统计学意义。结论:艾叶挥发油可降低AR大鼠血清中IL-4、IL-5和IgE含量,减轻鼻粘膜变应性炎症。     

"Bystander" amplification of PBMC cytokine responses to seasonal allergen in polysensitized atopic children

BACKGROUND: Atopic children show increased expression and production of the Th2-associated cytokines IL-4, IL-5, IL-13, and IL-9 from PBMCs after stimulation with allergen, but it has previously not been clearly determined whether the Th2-cytokine production is restricted to the inhalant allergen the child is sensitized to, and whether perennial or seasonal allergens induce different cytokine responses. Our purpose was to determine whether in vitro Th2 cytokine production is specific to the sensitizing allergen, and to compare the cytokine responses to a perennial and a seasonal allergen in monosensitized and polysensitized children. METHODS: Using semiquantitative RT-PCR, we analyzed the expression of the cytokines IL-4, IL-5, IL-13, IL-9, IL-10, and IFN-gamma after stimulation of PBMCs with house-dust-mite (HDM) or ryegrass allergen. The cells were sampled from groups of 6-year-old children sensitized to either HDM (n=20) or ryegrass (n=24), or to both allergens (n=20), as well as from a nonatopic group (n=20). RESULTS: After stimulation with HDM allergen, PBMCs from children sensitized only to HDM expressed increased mRNA levels of the Th2 cytokines, but not of IL-10 and IFN-gamma, whereas ryegrass stimulation did not result in increased cytokine expression. PBMCs from children sensitized to HDM and ryegrass expressed increased Th2 cytokines after stimulation with either of the two allergens. In contrast, PBMCs from children sensitized only to ryegrass did not express increased levels after stimulation with either of the allergens. CONCLUSIONS: The expression of Th2 cytokines after in vitro stimulation of PBMCs from atopic children is specific to the sensitizing allergen, indicating that atopic status per se does not affect the type of T-cell response. In addition, T cells specific to seasonal allergens circulate in the blood out of season only if the child is concomitantly sensitized to a perennial allergen.     

胰岛素对大鼠肝细胞损伤的保护及其抗炎机制

目的观察胰岛素对大鼠继发性肝细胞损伤是否具有保护作用并探讨其机制。方法胶原酶原位灌流分离大鼠枯否细胞(Kupffercell,KC)及肝细胞并原代培养,分别用脂多糖(LPS)及胰岛素处理KC4h,ELISA法检测KC培养上清液中肿瘤坏死因子-α(TNF-α)和白介素-10(IL-10)水平;将不同处理的KC培养上清液分别作用肝细胞,12h后检测肝细胞损伤及存活率,并给予TNF-α单克隆抗体(TNF-α-mAb)阻断TNF-α,观察胰岛素对肝细胞作用的变化。结果(1)LPS处理的KC培养上清液可引起肝细胞损伤,使丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)增高;胰岛素可减轻上述肝细胞损伤(ALT活性降低30.1%,AST活性降低21.2%)(P<0.01,n=8),提高肝细胞存活率(P<0.01,n=8);(2)胰岛素降低KC培养上清液中的TNF-α水平(P<0.01,n=8),同时增加了IL-10的水平(P<0.05,n=8);(3)LPS激活的KC培养上清中加入TNF-α-mAb(中和TNF-α),亦可减轻肝细胞损伤,而此时胰岛素对肝细胞的保护效应未见叠加增强。结论首次发现胰岛素可通过抑制KC分泌促炎性细胞因子TNF-α,同时促进其释放抗炎性细胞因子IL-10,从而减轻肝细胞损伤,促进肝细胞存活。     

Increased IL-2, IL-4 and interferon-gamma (IFN-gamma) in steroid-sensitive nephrotic syndrome.

We investigated the production of cytokines by peripheral blood mononuclear cells (PBMC) and serum cytokine concentrations in children with steroid-sensitive idiopathic nephrotic syndrome (SSNS). PBMC from patients off treatment were collected during remission and relapse and cultured in medium alone or stimulated with calcium ionophore plus phorbol myristate acetate. Control PBMC were taken from healthy age-matched children. IL-2 was measured by bioassay, IL-4 by immunoradiometric assay, and IL-8 and IFN-gamma by ELISA. After 24 h culture without stimulation, IL-2, IL-4 and IFN-gamma were not detectable in the supernatant in any of the children. After stimulation, the supernatant concentrations of IL-2 (median 172 U/ml at 24 h) and IL-4 (160 pg/ml at 24 h; 210 pg/ml at 72 h) were significantly increased in relapse compared with remission (IL-2 37 U/ml; IL-4 65 pg/ml and 60 pg/ml) and controls (IL-2 69 U/ml; IL-4 40 pg/ml and 40 pg/ml) (P < 0.05). The concentration of IFN-gamma was not significantly increased in relapse compared with remission and controls (600, 325, and 145 U/ml, respectively, at 72 h). IL-8 concentrations were similar in relapse, remission and controls with stimulation (median 32, 40 and 40 ng/ml, respectively) and without (30, 17 and 10 ng/ml). IL-2 was not detectable in serum, but IL-4, IL-8 and IFN-gamma were measurable in about half the patients, both in relapse and remission, though were virtually undetectable in controls. We conclude that relapse of SSNS in children is associated with T lymphocyte activation with release of IL-2, IL-4 and IFN-gamma.     

Evaluation of Th1/Th2 ratio and cytokine production profile during acute exacerbation and convalescence in asthmatic children.

BACKGROUND: Th2-type cytokines, such as IL-4 and IL-5, are generally believed to play an important role in the pathogenesis of allergic asthma. In contrast, Th1-type cytokine, especially interferon (IFN)-gamma, is thought to have a downregulatory effect on Th2 immune response cells. Thus, the imbalance of Th1 and Th2 cells may be a key factor in relation to disease severity. OBJECTIVE: The aim of this study is to examine the changes in the Th1/Th2 ratios and cytokine production profiles of asthmatic children at acute attacks and convalescent stages. METHODS: Twelve asthmatic patients were included in this study. The percentages of IFN-gamma- or IL-4-producing cells were determined with a flow-cytometric method of intracellular protein detection. Fresh whole blood obtained from normal controls and patients at two stages was stimulated with brefeldin A, phorbol myristate acetate, and ionomycin for 4 hours. Cells were fixed and stained intracellularly with fluorescein isothiocyanate- or phycoerythrin-conjugated antibody specific to each cytokine in combination with anti-CD4. ELISA assays were applied to measure cytokine concentrations of supernatant from peripheral blood mononuclear cells (PBMC) activated with phorbol myristate acetate and ionomycin for 24 hours. RESULTS: Among CD4+ cells, the percentage of IL-4+ cells decreased significantly from 8.18 +/- 4.77% at acute attacks to 4.18 +/- 1.26% (P < .020) at convalescence. The percentage of IFN-gamma+ cells also decreased from 13.49 +/- 4.21% to 9.03 +/- 5.42% (P < .052). The Th1/Th2 ratios of patients at the two stages were similar, and both were lower than the normal controls. Significantly higher IL-5 and lower IFN-gamma production were detected from activated PBMC of asthmatic patients than normal controls. CONCLUSIONS: The decrease of IFN-gamma+ and IL-4+ cells detected at the single-cell level may explain the potential mechanism of convalescence from acute asthma attacks. High Th1/Th2 ratio and low IL-5 production from the PBMC of normal controls support the idea of a biased Th2 immune response in asthmatic patients.     

CTLA4-Ig在小鼠病毒性心肌炎中作用机制的研究

目的 探讨细胞毒T淋巴细胞相关抗原4融合蛋白(CTLA4-Ig)对柯萨舒B3(CVB3)病毒性心肌炎(VMC)小鼠死亡率、心肌病理改变与病毒滴度、CTLA4蛋白表达及Th1/Th2平衡的影响.方法 将106只BALB/c小鼠随机分为CTLA4-Ig组16只、病毒组40只、IgG组40只及正常对照组10只,于接种病毒后第7天处死所有小鼠,采用光镜检查心肌病理变化和实时荧光定量PCR(RQ-PCR)检测心肌组织中CVB3 mRNA的表达,免疫组化法检测CTLA4蛋白的表达,取外周血应用酶联免疫吸附法(ELISA)检测血清IL-2、IL-4及IFN-γ的含最.结果 CTLA4-Ig组小鼠死亡率、心肌病理积分、心肌CVB3 mRNA均低于病毒对照组(P均<0.05);CTLA4-Ig组小鼠心肌组织中CTLA4表达较病毒组明显增加(P<0.05);病毒组小鼠血清IFN-γ水平明显高于正常对照组小鼠(P<0.01),IL-4水平明显低于正常对照组小鼠(P<0.01),两组小鼠间IL-2的水平差异无统计学意义(P>0.05).CTLA4-Ig组小鼠血清IL-4水平明显高于病毒组及IgG组(P均<0.01),IFN-γ水平低于病毒组及IgG组(P均<0.01),三组小鼠血清IL-2水平的差异无统计学意义(P均>0.05).结论 CTLA4-Ig可减轻VMC小鼠心肌炎症,降低心肌病毒滴度及死亡率,其机制可能与阻断T细胞活化的共刺激信号,使Th2反应增强有关.     

Adult Still's disease reflects a Th2 rather than a Th1 cytokine profile

Adult Still's disease (ASD) is a chronic multisystemic disease. Extraordinarily high serum levels of IL-18 in ASD patients have been described, whereas the mechanism remains to be clarified. This study aimed to evaluate proinflammatory cytokines and to consider their pathological roles. In patients with rheumatic diseases (n = 151), blood samples were taken at the active phase and the serum levels of IL-18 and other proinflammatory cytokines were measured by ELISA. The extra-high levels of IL-18 were confirmed selectively in ASD patients (n = 10). In the active phase of ASD patients, the levels of IL-6 were elevated accordingly, but IL-1beta and TNF-alpha were undetectable. As to Th1-Th2 cytokines, the levels of IL-4 and IL-13, but not INF-gamma, IL-12, or IL-2, were elevated in all ASD patients examined. Moreover, the serum levels of IL-18 showed a good correlation with those of IL-4, suggesting that ASD reflects a Th2 rather than a Th1 cytokine profile.